Microplastics are ubiquitous in the environment, with high concentrations being detected now also in river corridors and sediments globally. Whilst there has been increasing field evidence of microplastics accumulation in the guts and tissues of freshwater and marine aquatic species, the uptake mechanisms of microplastics into freshwater food webs, and the physical and geological controls on pathway-specific exposures to microplastics, are not well understood. This knowledge gap is hampering the assessment of exposure risks, and potential ecotoxicological and public health impacts from microplastics.This review provides a comprehensive synthesis of key research challenges in analysing the environmental fate and transport of microplastics in freshwater ecosystems, including the identification of hydrological, sedimentological and particle property controls on microplastic accumulation in aquatic ecosystems. This mechanistic analysis outlines the dominant pathways for exposure to microplastics in freshwater ecosystems and identifies potentially critical uptake mechanisms and entry pathways for microplastics and associated contaminants into aquatic food webs as well as their risk to accumulate and biomagnify.We identify seven key research challenges that, if overcome, will permit the advancement beyond current conceptual limitations and provide the mechanistic process understanding required to assess microplastic exposure, uptake, hazard, and overall risk to aquatic systems and humans, and provide key insights into the priority impact pathways in freshwater ecosystems to support environmental management decision making.

International audience | Microplastics are ubiquitous in the environment, with high concentrations being detected now also in river corridors and sediments globally. Whilst there has been increasing field evidence of microplastics accumulation in the guts and tissues of freshwater and marine aquatic species, the uptake mechanisms of microplastics into freshwater food webs, and the physical and geological controls on pathway-specific exposures to microplastics, are not well understood. This knowledge gap is hampering the assessment of exposure risks, and potential ecotoxicological and public health impacts from microplastics.This review provides a comprehensive synthesis of key research challenges in analysing the envi- ronmental fate and transport of microplastics in freshwater ecosystems, including the identification of hydrological, sedimentological and particle property controls on microplastic accumulation in aquatic ecosystems. This mechanistic analysis outlines the dominant pathways for exposure to microplastics in freshwater ecosystems and identifies potentially critical uptake mechanisms and entry pathways for microplastics and associated contaminants into aquatic food webs as well as their risk to accumulate and biomagnify.We identify seven key research challenges that, if overcome, will permit the advancement beyond current conceptual limitations and provide the mechanistic process understanding required to assess microplastic exposure, uptake, hazard, and overall risk to aquatic systems and humans, and provide key insights into the priority impact pathways in freshwater ecosystems to support environmental man- agement decision making.

Antarctic melt streams are important ecosystems that increasingly face contaminant pressures from anthropogenic sources. Metal contaminants are often reported in the limno-terrestrial environment but their speciation is not well characterised, making environmental risk assessments difficult. This paper characterises labile metal concentrations in five melt streams and three shallow lakes around the Casey and Wilkes research stations in East Antarctica using chemical extracts and field deployments of diffusive gradients in thin-film (DGT) samplers. An acute toxicity test with field-collected Ceratadon purpeus and taxonomic identification of diatoms in melt streams were used to infer environmental risk. Copper and zinc were the most labile metals in the melt streams. DGT-labile copper concentrations were up to 3 μg Cu L⁻¹ in melt-stream waters but not labile below the sediment-water interface. DGT-labile zinc concentrations were consistent above and below the sediment-water interface at concentrations up to 14 μg Zn L⁻¹ in four streams, but one stream showed evidence of zinc mineralisation in the sediment with a flux to overlying and pore waters attributed to the reductive dissolution of iron and manganese oxides. Other metals, such as chromium, nickel, and lead were acid-extractable from the sediments, but not labile in pore waters or overlying waters. All streams had unique compositions of freshwater diatoms, but one had particularly reduced diversity and richness, which correlated to metal contamination and sediment physico-chemical properties such as a finer particle size. In laboratory bioassays with field-collected samples of the Antarctic moss C. purpeus, there was no change in photosynthetic efficiency following 28-d exposure to 700, 900, 1060, or 530 μg L⁻¹ of cadmium, copper, nickel, and zinc, respectively. This study shows that microorganisms such as diatoms may be at greater risk from contaminants than mosses, and highlights the importance of geochemical factors controlling metal lability.

With a rule published on 6 January 2021, the US Environmental Protection Agency (EPA) considers for the first time sub-threshold responses, abandoning the use of default dose-response models. This may affect worldwide scientific research, in terms of research design and methodology, and regulatory actions in China and other countries.

With a rule published on 6 January 2021, the US Environmental Protection Agency (EPA) considers for the first time sub-threshold responses, abandoning the use of default dose-response models. This may affect worldwide scientific research, in terms of research design and methodology, and regulatory actions in China and other countries. | This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. E.A. acknowledges multi-year support from The Startup Foundation for Introducing Talent of Nanjing University of Information Science and Technology, Nanjing, China (No. 003080 to E.A.). E.J.C. acknowledges longtime support from the U.S. Air Force (AFOSR FA9550-13-1-0047) and ExxonMobil Foundation (S18200000000256). The sponsors were not involved in the study design; the collection, analysis or interpretation of the data; the preparation of the manuscript or the decision where to submit the manuscript for publication. | Peer reviewed

The decline of shark populations in the world ocean is affecting ecosystem structure and function in an unpredictable way and new ecological information is today needed to better understand the role of sharks in their habitats. In particular, the characterization of foraging patterns is crucial to understand and foresee the evolution of dynamics between sharks and their prey. Many shark species use the mesopelagic area as a major foraging ground but the degree to which different pelagic sharks rely on this habitat remains overlooked. In order to depict the vertical dimension of their trophic ecology, we used mercury stable isotopes in the muscle of three pelagic shark species (the blue shark Prionace glauca, the shortfin mako shark Isurus oxyrinchus and the smooth hammerhead shark Sphyrna zygaena) from the northeastern Pacific region. The Δ¹⁹⁹Hg values, ranging from 1.40 to 2.13‰ in sharks, suggested a diet mostly based on mesopelagic prey in oceanic habitats. We additionally used carbon and nitrogen stable isotopes (δ¹³C, δ¹⁵N) alone or in combination with Δ¹⁹⁹Hg values, to assess resource partitioning between the three shark species. Adding Δ¹⁹⁹Hg resulted in a decrease in trophic overlap estimates compared to those based on δ¹³C/δ¹⁵N alone, demonstrating that multi-isotope modeling is needed for accurate trophic description of the three species. Mainly, it reveals that they forage at different average depths and that resource partitioning is mostly expressed through the vertical dimension within pelagic shark assemblages. Concomitantly, muscle total mercury concentration (THg) differed between species and increased with feeding depth. Overall, this study highlights the key role of the mesopelagic zone for shark species foraging among important depth gradients and reports new ecological information on trophic competition using mercury isotopes. It also suggests that foraging depth may play a pivotal role in the differences between muscle THg from co-occurring high trophic level shark species.

The decline of shark populations in the world ocean is affecting ecosystem structure and function in an unpredictable way and new ecological information is today needed to better understand shark roles in their habitats. In particular, the characterization of foraging patterns is crucial to understand and foresee the evolution of dynamics between sharks and their prey. Many shark species use the mesopelagic area as a major foraging ground but the degree to which different pelagic sharks rely on this habitat remains overlooked. In order to depict the vertical dimension of their trophic ecology, we used mercury stable isotopes in the muscle of three pelagic shark species (the blue shark Prionace glauca, the shortfin mako shark Isurus oxyrinchus and the smooth hammerhead shark Sphyrna zygaena) from the northeastern Pacific region. The Δ199Hg values, ranging from 1.40 to 2.13 ‰ in sharks, suggested a diet mostly based on mesopelagic prey in oceanic habitats. We additionally used carbon and nitrogen stable isotopes (δ13C, δ15N) alone or in combination with Δ199Hg values, to assess resource partitioning between the three shark species. Combining Δ199Hg resulted in a decrease in trophic overlap estimates compared to δ13C/δ15N alone, demonstrating that multi-isotope modeling is needed for accurate trophic description of the three species. Mainly, it reveals that they forage at different average depths and that resource partitioning is mostly expressed through the vertical dimension within pelagic shark assemblages. Concomitantly, muscle total mercury concentration (THg) differed between species and increased with feeding depth. Overall, this study highlights the key role of the mesopelagic zone for shark species foraging among important depth gradients and reports new ecological information on trophic competition using mercury isotopes. It also suggests that foraging depth may play a pivotal role in the differences between muscle THg from co-occurring high trophic level shark species.

Plastic residues have become a serious environmental problem in areas where agricultural plastic film are used intensively. Although numerous of studies have been done to assess its impacts on soil quality and crop yields, the understanding of meso-plastic particles effects on plant is still limited. In this study, low density polyethylene (PE) and biodegradable plastic (Bio) mulch film were selected to study the effects of meso-plastic debris on soybean germination and plant growth with the accumulation levels of 0%, 0.1%, 0.5% and 1% in soil (w: w, size ranging 0.5–2 cm) by a pot experiment under field condition. Results showed that the germination viability of soybean seeds was reduced to 82.39%, 39.44% and 26.06% in the treatments with 0.1%, 0.5% and 1% added plastic debris compared to the control (CK), respectively, suggesting that plastic residues in soil inhibit the viability of soybean seed germination. The plastic debris had a significant negative effect on plant height and culm diameter during the entire growth stage of soybean. Similarly, the leaf area at harvest was reduced by 1.97%, 6.86% and 11.53% compared to the CK in the treatments with 0.1%, 0.5% and 1% plastic debris addition, respectively. In addition, the total plant biomass under plastic addition was reduced in both the flowering and harvesting stages, compared to the CK. For the different type of plastic residues, plant height, leaf area and root/shoot ratio at group PE were significantly lower than those of groups treated by Bio. In conclusion, PE debris had a greater negative effects on plant height, culm diameter, leaf area and root/shoot ratio while Bio debris mainly showed the adverse effects on germination viability and root biomass especially at the flowering stage. Therefore, further research is required to elaborate plastic particles’ effects on different stages of crops and soil quality.

Plastic residues have become a serious environmental problem in areas where agricultural plastic film are used intensively. Although numerous of studies have been done to assess its impacts on soil quality and crop yields, the understanding of meso-plastic particles effects on plant is still limited. In this study, low density polyethylene (PE) and biodegradable plastic (Bio) mulch film were selected to study the effects of meso-plastic debris on soybean germination and plant growth with the accumulation levels of 0%, 0.1%, 0.5% and 1% in soil (w: w, size ranging 0.5–2 cm) by a pot experiment under field condition. Results showed that the germination viability of soybean seeds was reduced to 82.39%, 39.44% and 26.06% in the treatments with 0.1%, 0.5% and 1% added plastic debris compared to the control (CK), respectively, suggesting that plastic residues in soil inhibit the viability of soybean seed germination. The plastic debris had a significant negative effect on plant height and culm diameter during the entire growth stage of soybean. Similarly, the leaf area at harvest was reduced by 1.97%, 6.86% and 11.53% compared to the CK in the treatments with 0.1%, 0.5% and 1% plastic debris addition, respectively. In addition, the total plant biomass under plastic addition was reduced in both the flowering and harvesting stages, compared to the CK. For the different type of plastic residues, plant height, leaf area and root/shoot ratio at group PE were significantly lower than those of groups treated by Bio. In conclusion, PE debris had a greater negative effects on plant height, culm diameter, leaf area and root/shoot ratio while Bio debris mainly showed the adverse effects on germination viability and root biomass especially at the flowering stage. Therefore, further research is required to elaborate plastic particles’ effects on different stages of crops and soil quality.

During the last years, ingestion of microplastics (MPs) has been quantified in marine species both with an ecological and commercial interest at sea and under experimental conditions, highlighting the importance to assess MP ingestion in commercially and aquaculture important species such as gilthead seabream (Sparus aurata) fish. In order to study the ingestion of MPs in a commercially valuable species, gilthead seabreams were exposed to an enriched diet with virgin and weathered low-density polyethylene (LDPE) pellets for three months followed by a detoxification period of one month of no exposure to MP enriched diets. Our results indicate that MP ingestion in these fishes increased with exposure time, and differences were found between treatments, showing the highest ingestion values after three months of exposure to MP enriched diets and in the weathered treatment. However, after one month of detoxification, no MPs were found in the gastrointestinal tracts of fish, reflecting no long-term retention of MPs in Sparus aurata digestive system. According to results from this study, exposure of fish to MP enriched diets does not affect fish size neither the Fulton’s condition index as both parameters increased with time in all treatments (control, virgin and weathered). Both carbon and nitrogen isotopic signatures decreased with fish size in all treatments which could be related to an increase of nitrogen deposition efficiency in fish muscle with a high protein assimilation during the first months of Sparus aurata.

During the last years, ingestion of microplastics (MPs) has been quantified in marine species both with an ecological and commercial interest at sea and under experimental conditions, highlighting the importance to assess MP ingestion in commercially and aquaculture important species such as gilthead seabream (Sparus aurata) fish. In order to study the ingestion of MPs in a commercially valuable species, gilthead seabreams were exposed to an enriched diet with virgin and weathered low-density polyethylene (LDPE) pellets for three months followed by a detoxification period of one month of no exposure to MP enriched diets. Our results indicate that MP ingestion in these fishes increased with exposure time, and differences were found between treatments, showing the highest ingestion values after three months of exposure to MP enriched diets and in the weathered treatment. However, after one month of detoxification, no MPs were found in the gastrointestinal tracts of fish, reflecting no long-term retention of MPs in Sparus aurata digestive system. According to results from this study, exposure of fish to MP enriched diets does not affect fish size neither the Fulton’s condition index as both parameters increased with time in all treatments (control, virgin and weathered). Both carbon and nitrogen isotopic signatures decreased with fish size in all treatments which could be related to an increase of nitrogen deposition efficiency in fish muscle with a high protein assimilation during the first months of Sparus aurata. © 2021 Elsevier Ltd. All rights reserved. | This study was funded by the Aquaculture derived plastics: Impacts and effects in marine food webs (ACUIPLASTIC) project from the National Program of Investigation, Development and Innovation orientated towards challenges of society from the Spanish Government (grant number CTM 2017-88332-R). The authors acknowledge financial support from the Spanish State Research Agency (AEI) and the Ministry of Science, Innovation and Universities (MICIU) to support the Thematic Network of Excellence (NET4SEA) on emerging contaminants in marine settings (CTM 2017-90890-REDT, MICIU/AEI/FEDER, EU). F. Alarcón from the Biology and Geology department of the Escuela Polítecnica Superior of the University of Almería for MPs enriched diet preparation. Authors want to acknowledge that M. Compa was supported by an FPI Fellowship from Conselleria d’Innovació, Recerca i Turisme of the regional Government of the Balearic Islands co-financed by the European Social Fund as part of the FSE 2014–2020 operational program. E. Álvarez was supported by a Personal Técnico de Apoyo contract (PTA 2015-10829-I) funded by the Spanish Ministry of Economy and Competitiveness. Authors want to thank the four anonymous reviewers for the constructive revision. | Peer reviewed

Two different methodologies were combined to evaluate the risks that antibiotics can pose in the environment; i) an effect-based methodology based on microbial growth inhibition and ii) an analytical method based on liquid-chromatography coupled to mass spectrometry (LC-MS). The first approach was adapted and validated for the screening of four antibiotic families, specifically macrolides/β-lactams, quinolones, sulfonamides and tetracyclines. The LC-MS method was applied for the identification and quantification of target antibiotics; then, the obtained results were combined with ecotoxicological data from literature to determine the environmental risk. The two methodologies were used for the analysis of antibiotics in water samples (wastewater, river water and seawater) and biofluids (fish plasma and mollusk hemolymph) in two monitoring campaigns undertaken in the Ebro Delta and Mar Menor Lagoon (both in the Mediterranean coast of Spain). Both approaches highlighted macrolides (azithromycin) and quinolones (ciprofloxacin and ofloxacin) as the main antibiotics in wastewater treatment plant (WWTP) effluents with potential risk for the environment. However, no risk for the aquatic life was identified in the river, lagoon and seawater as antibiotic levels were much lower than those in WWTP effluents. Fish from Ebro River were the organisms presenting the highest antibiotic concentration when compared with bivalves (mussels) from the Mediterranean Sea and gastropods (marine snails) from the Mar Menor Lagoon. The effect-based methodology successfully determined antibiotic risk in wastewater, but its applicability was less clear in environmental waters such as seawater, due to its high detection limits. Improving sample preconcentration could increase the method sensibility. Overall, combination of both methodologies provides comprehensive insights in antibiotic occurrence and risk associated in areas under study.

Two different methodologies were combined to evaluate the risks that antibiotics can pose in the environment; i) an effect-based methodology based on microbial growth inhibition and ii) an analytical method based on liquid-chromatography coupled to mass spectrometry (LC-MS). The first approach was adapted and validated for the screening of four antibiotic families, specifically macrolides/β-lactams, quinolones, sulfonamides and tetracyclines. The LC-MS method was applied for the identification and quantification of target antibiotics; then, the obtained results were combined with ecotoxicological data from literature to determine the environmental risk. The two methodologies were used for the analysis of antibiotics in water samples (wastewater, river water and seawater) and biofluids (fish plasma and mollusk hemolymph) in two monitoring campaigns undertaken in the Ebro Delta and Mar Menor Lagoon (both in the Mediterranean coast of Spain). Both approaches highlighted macrolides (azithromycin) and quinolones (ciprofloxacin and ofloxacin) as the main antibiotics in wastewater treatment plant (WWTP) effluents with potential risk for the environment. However, no risk for the aquatic life was identified in the river, lagoon and seawater as antibiotic levels were much lower than those in WWTP effluents. Fish from Ebro River were the organisms presenting the highest antibiotic concentration when compared with bivalves (mussels) from the Mediterranean Sea and gastropods (marine snails) from the Mar Menor Lagoon. The effect-based methodology successfully determined antibiotic risk in wastewater, but its applicability was less clear in environmental waters such as seawater, due to its high detection limits. Improving sample preconcentration could increase the method sensibility. Overall, combination of both methodologies provides comprehensive insights in antibiotic occurrence and risk associated in areas under study. | This work has received funding from the Spanish Ministry of Economy and Competitiveness through the PLAS-MED (CTM2017-89701-C3-2-R) project. Authors acknowledge the support from the Economy and Knowledge Department of the Catalan Government through Consolidated Research Group (ICRA-ENV 2017 SGR 1124 and 2017-SGR-1404-Water and Soil Quality Unit). The authors acknowledge the IEO & CSIC staff for their help in the sampling campaigns. Albert Serra-Compte acknowledges the FI-DGR research fellowship from the Catalan Government (2018FI_B2_00170) and the Nereus Cost Action grant. Sara Rodriguez-Mozaz acknowledges the Ramon y Cajal program (RYC-2014-16707) and Diana Álvarez-Muñoz the support of the project XENOMETABOLOMIC (CTM2015-73179-JIN) (AEI/FEDER/UE). Authors kindly acknowledge Generalitat DGPAM (Fisheries Department). The authors also acknowledge support from CERCA Programme/Generalitat de Catalunya. | Peer reviewed

Marine canopies formed by seagrass and other coastal vegetated ecosystems could act as sinks of microplastics for being efficient particle traps. Here we investigated for the first time the occurrence of microplastic retention by marine canopies in a hydraulic flume under unidirectional flow velocities from 2 to 30 cm s⁻¹. We used as model canopy-forming species the seagrass Zostera marina with four canopy shoot density (0, 50, 100, 200 shoots m⁻²), and we used as microplastic particles industrial pristine pellets with specific densities from 0.90 to 1.34 g cm⁻³ (polypropylene PP; polystyrene PS; polyamide 6 PA; and polyethylene terephthalate PET). Overall, microplastics particles transported with the flow were retained in the seagrass canopies but not in bare sand. While seagrass canopies retained floating microplastics (PP) only at low velocities (<12 cm s⁻¹) due to a barrier created by the canopy touching the water surface, the retention of sinking particles (PS, PA, PET) occurred across a wider range of flow velocities. Our simulations revealed that less dense sinking particles (PS) might escape from the canopy at high velocities, while denser sinking particles can be trapped in scouring areas created by erosive processes around the eelgrass shoots. Our results show that marine canopies might act as potential barriers or sinks for microplastics at certain bio-physical conditions, with the probability of retention generally increasing with the seagrass shoot density and polymer specific density and decreasing with the flow velocity. We conclude that seagrass meadows, and other aquatic canopy-forming ecosystems, should be prioritized habitats in assessment of microplastic exposure and impact on coastal areas since they may accumulate high concentration of microplastic particles that could affect associated fauna.

Marine canopies formed by seagrass and other coastal vegetated ecosystems could act as sinks of microplastics for being efficient particle traps. Here we investigated for the first time the occurrence of microplastic retention by marine canopies in a hydraulic flume under unidirectional flow velocities from 2 to 30 cm s−1. We used as model canopy-forming species the seagrass Zostera marina with four canopy shoot density (0, 50, 100, 200 shoots m−2), and we used as microplastic particles industrial pristine pellets with specific densities from 0.90 to 1.34 g cm−3 (polypropylene PP; polystyrene PS; polyamide 6 PA; and polyethylene terephthalate PET). Overall, microplastics particles transported with the flow were retained in the seagrass canopies but not in bare sand. While seagrass canopies retained floating microplastics (PP) only at low velocities (<12 cm s−1) due to a barrier created by the canopy touching the water surface, the retention of sinking particles (PS, PA, PET) occurred across a wider range of flow velocities. Our simulations revealed that less dense sinking particles (PS) might escape from the canopy at high velocities, while denser sinking particles can be trapped in scouring areas created by erosive processes around the eelgrass shoots. Our results show that marine canopies might act as potential barriers or sinks for microplastics at certain bio-physical conditions, with the probability of retention generally increasing with the seagrass shoot density and polymer specific density and decreasing with the flow velocity. We conclude that seagrass meadows, and other aquatic canopy-forming ecosystems, should be prioritized habitats in assessment of microplastic exposure and impact on coastal areas since they may accumulate high concentration of microplastic particles that could affect associated fauna. | publishedVersion

Marine canopies formed by seagrass and other coastal vegetated ecosystems could act as sinks of microplastics for being efficient particle traps. Here we investigated for the first time the occurrence of microplastic retention by marine canopies in a hydraulic flume under unidirectional flow velocities from 2 to 30 cm s-1. We used as model canopy-forming species the seagrass Zostera marina with four canopy shoot density (0, 50, 100, 200 shoots m-2), and we used as microplastic particles industrial pristine pellets with specific densities from 0.90 to 1.34 g cm-3 (polypropylene PP; polystyrene PS; polyamide 6 PA; and polyethylene terephthalate PET). Overall, microplastics particles transported with the flow were retained in the seagrass canopies but not in bare sand. While seagrass canopies retained floating microplastics (PP) only at low velocities (<12 cm s-1) due to a barrier created by the canopy touching the water surface, the retention of sinking particles (PS, PA, PET) occurred across a wider range of flow velocities. Our simulations revealed that less dense sinking particles (PS) might escape from the canopy at high velocities, while denser sinking particles can be trapped in scouring areas created by erosive processes around the eelgrass shoots. Our results show that marine canopies might act as potential barriers or sinks for microplastics at certain bio-physical conditions, with the probability of retention generally increasing with the seagrass shoot density and polymer specific density and decreasing with the flow velocity. We conclude that seagrass meadows, and other aquatic canopy-forming ecosystems, should be prioritized habitats in assessment of microplastic exposure and impact on coastal areas since they may accumulate high concentration of microplastic particles that could affect associated fauna. | FCT: UIDB/04326/2020/ SFRH/BPD/119344/2016 | info:eu-repo/semantics/publishedVersion

Several studies highlighted the ubiquitous presence of ibuprofen and aluminum in the aquatic environment around the world and demonstrated their potential to induce embryotoxic and teratogenic defects on aquatic species individually. Although studies that evaluate developmental alterations induced by mixtures of these pollutants are scarce; and, since environmental contamination presented in the form of a mixture of toxicants with different chemical properties and toxicity mechanisms capable of generating interactions; the objective of this study was to evaluate the developmental defects, teratogenic alterations, and oxidative stress induced by individual forms and the mixture of ibuprofen (IBU) and aluminum (Al) on zebrafish embryos. Oocytes exposed to environmentally relevant concentrations of IBU (0.1–20 μg L-1) and Al (0.01–8 mg L-1) and one binary mixture. The LC50 and EC50 were obtained to calculate the teratogenic index (TI). The IBU LC50, EC50, and TI were 8.06 μg L-1, 2.85 μg L-1 and 2.82. In contrast, Al LC50 was 5.0 mg L-1with an EC50 of 3.58 mg L-1 and TI of 1.39. The main alterations observed for individual compounds were hatching alterations, head malformation, skeletal deformities, hypopigmentation, pericardial edema, and heart rate impairment. The mixture also showed significant delays to embryonic development. Moreover, oxidative stress biomarkers of cellular oxidation and antioxidant defenses at 72 and 96 hpf significantly increased. Results show that environmentally relevant concentrations of ibuprofen (IBU), aluminum (Al), and their mixture promote a series of developmental defects, teratogenic effects, and oxidative disruption on D. rerio embryos, and the interaction of both substances altered the response. In conclusion, morphological and biochemical tests are suitable tools for assessing the health risk of aquatic wildlife by exposure to individual and mixed pollutants in freshwater bodies.

Several studies highlighted the ubiquitous presence of ibuprofen and aluminum in the aquatic environment around the world and demonstrated their potential to induce embryotoxic and teratogenic defects on aquatic species individually. Although studies that evaluate developmental alterations induced by mixtures of these pollutants are scarce; and, since environmental contamination presented in the form of a mixture of toxicants with different chemical properties and toxicity mechanisms capable of generating interactions; the objective of this study was to evaluate the developmental defects, teratogenic alterations, and oxidative stress induced by individual forms and the mixture of ibuprofen (IBU) and aluminum (Al) on zebrafish embryos. Oocytes exposed to environmentally relevant concentrations of IBU (0.1–20 μg L-1) and Al (0.01–8 mg L-1) and one binary mixture. The LC50 and EC50 were obtained to calculate the teratogenic index (TI). The IBU LC50, EC50, and TI were 8.06 μg L-1, 2.85 μg L-1 and 2.82. In contrast, Al LC50 was 5.0 mg L-1with an EC50 of 3.58 mg L-1 and TI of 1.39. The main alterations observed for individual compounds were hatching alterations, head malformation, skeletal deformities, hypopigmentation, pericardial edema, and heart rate impairment. The mixture also showed significant delays to embryonic development. Moreover, oxidative stress biomarkers of cellular oxidation and antioxidant defenses at 72 and 96 hpf significantly increased. Results show that environmentally relevant concentrations of ibuprofen (IBU), aluminum (Al), and their mixture promote a series of developmental defects, teratogenic effects, and oxidative disruption on D. rerio embryos, and the interaction of both substances altered the response. In conclusion, morphological and biochemical tests are suitable tools for assessing the health risk of aquatic wildlife by exposure to individual and mixed pollutants in freshwater bodies. | This study was made possible by financial support from the Consejo Nacional de Ciencia y Tecnología (CONACyT, Project 300727). | Peer reviewed

Chronic toxicity of copper (Cu) at sublethal levels is associated with ionoregulatory disturbance and oxidative stress. These factors were considered in a toxicokinetic-toxicodynamic model in the present study. The ionoregulatory disturbance was evaluated by the activity of the Na⁺/K⁺-ATPase enzyme (NKA), while oxidative stress was presented by lipid peroxidation (LPO) and glutathione-S-transferase (GST) activity. NKA activity was related to the binding of Cu²⁺ and Na ⁺ to NKA. LPO and GST activity were linked with the simulated concentration of unbound Cu. The model was calibrated using previously reported data and empirical data generated when zebra mussels were exposed to Cu. The model clearly demonstrated that Cu might inhibit NKA activity by reducing the number of functional pump sites and the limited Cu-bound NKA turnover rate. An ordinary differential equation was used to describe the relationship between the simulated concentration of unbound Cu and LPO/GST activity. Although this method could not explain the fluctuations in these biomarkers during the experiment, the measurements were within the confidence interval of estimations. Model simulation consistently shows non-significant differences in LPO and GST activity at two exposure levels, similar to the empirical observation.

10 pages, 2 figures, 3 tables | Chronic toxicity of copper (Cu) at sublethal levels is associated with ionoregulatory disturbance and oxidative stress. These factors were considered in a toxicokinetic-toxicodynamic model in the present study. The ionoregulatory disturbance was evaluated by the activity of the Na+/K+-ATPase enzyme (NKA), while oxidative stress was presented by lipid peroxidation (LPO) and glutathione-S-transferase (GST) activity. NKA activity was related to the binding of Cu2+ and Na + to NKA. LPO and GST activity were linked with the simulated concentration of unbound Cu. The model was calibrated using previously reported data and empirical data generated when zebra mussels were exposed to Cu. The model clearly demonstrated that Cu might inhibit NKA activity by reducing the number of functional pump sites and the limited Cu-bound NKA turnover rate. An ordinary differential equation was used to describe the relationship between the simulated concentration of unbound Cu and LPO/GST activity. Although this method could not explain the fluctuations in these biomarkers during the experiment, the measurements were within the confidence interval of estimations. Model simulation consistently shows non-significant differences in LPO and GST activity at two exposure levels, similar to the empirical observation | This research was financed by the Deutsche Forschungsgemeinschaft (DFG), Germany (LE 3716/2-1) | Peer reviewed